When entering the 21st century, with the aging of the population, the morbidity of senile dementia is increasing year by year in the world. There are about 20 million people suffering from senile dementia around the world as reported, wherein there are about 5-7 million people in China. The number of people who died because of suffering from senile dementia is in the fourth place after heart disease, cancer and stroke. In China, the morbidity of dementia of people over 65 years old is 6-7%; the morbidity of dementia of people over 70 years old is 13-15%; the morbidity of dementia of people over 80 years old is 22-25%; unfortunately, there are no better methods to effectively treat and prevent the senile dementia.
The senile dementia comprises Alzheimer's disease (AD) and vascular dementia (VD), the modern medical science has not figured out the pathogenesis of AD, and it is currently considered to be a progressive and neuro-degenerative disease. However, the VD is various dementias resulted from neurocranium internal hemorrhage, multiple infarct, arteriosclerosis encephalopathy acute brain trauma, encephalic space-occupying lesion, alcoholic intoxication, brain nutritional deficiency and metabolic endocrine disorder.
No matter what results in the senile dementia, the early symptoms are all forgetfulness, and hypesthesia of computational power and language competence; the second phase is serious memory loss, direction loss and of few words; and the third phase is serious thinking capacity disorder and vague orientation. Therefore, for studying the prevention and treatment of the senile dementia, it is prime to prevent memory and intelligence decline, and the ginsenoside Rg3 has significant effects of improving memory and delaying the ageing, which is also one of objectives of the present invention.
At present, a large number of medical research materials prove that the cerebral cortex and hippocampal cholinergic neurons of people suffering from senile dementia are largely lost, the presynaptic specific cholinergic mark choline acetyl transferase (ChAT) of a cholinergic transfer medium acetyl choline (Ach) is remarkably decreased, and this is one of disease causes of the senile dementia, so how to increase the functions of a reticular formation ascending activating system of the brain stem and promote Ach synthesis is an effective method for preventing and treating the senile dementia. Ginsenoside Rg3 can effectively activate the functions of the reticular formation ascending activating system of the brain stem, promote the Ach synthesis, increase cerebral blood flow, improve microcirculation, and therefore can be used for preventing and treating the senile dementia, and this is also the second objective of the present invention.
At present, the medicines used for preventing and treating the dementia mainly include donepezil, rivastigmine, galantamine, piracetam, aniracetam, vinpocetine, nicergoline, dihydroergotoxine, huperaine A, ginkgo leaves agent, brain protein hydrolysate, citicoline, cholinesterase inhibitor, xanthine, nonsteroidal anti-inflammatory medicine, estrogen, etc. Although these medicines have a certain relieving effect, but side effects are great, limiting their general use.
Chinese medicines for treatment of senile dementia are as follows: firstly, on the aspect of treatment from the standpoint of liver and kidney, body essence and marrow supplementing prescriptions are mostly adopted, such as Radix Rehmanniae Preparata, Fructus Corni, Rhizoma Polygonati, Carapax et Plastrum Testudinis, Colla Corii Asini, Fructus Lycii, Cornu Cervi Pantotrichu, etc; secondly, on the aspect of treatment through phlegm, the prescriptions for invigorating spleen to eliminate dampness, eliminating phlegm, inducing resuscitation and restoring consciousness are mostly adopted, such as Radix Curcumae, Rhizoma Acori Tatarinowii, Radix Polygalae, Fructus Alpiniae Oxyphyllae, XINGSHEN JIAONANG and the like;
thirdly, on the aspect of treatment through QI-stagnation and blood stasis, the prescriptions for regulating QI, activating blood circulation, inducing resuscitation and restoring consciousness are mostly adopted, such as DANGGUI-SHAOYAO SAN, BUYANGHUANWU TANG, TONGMAIYIZHI DAN and the like.
However, these Chinese medicines needs to be administered by a herbalist doctor based on differential treatment that varies from person to person, and cannot be used by the patients alone as they are incapable of taking these Chinese medicines properly. The effective ingredient ginsenoside Rg3 extracted from natural plants of the present invention have high purity (80% or above 80%) and is unique in curative effect, a prepared preparation is convenient to use, patients can adjust the dosages according to self reactions by themselves, and the effect of the ginsenoside after transdermal absorption is moderate and lasting.
Panax ginseng C. A. Mey. is an araliaceae perennial herb and is regarded as the top grade and described as that the Radix Ginseng mainly reinforces internal organs, soothes mind and body tranquilization, calms spirits, relieves palpitation due to fear, improves eyesight, is uplifting, is beneficial to intelligence and has the effects of reducing body weight and prolonging life after being taken for a long time in the Chinese pharmaceutical ancient book in regard to traditional Chinese medicine <Shen Nong's Herbal Classic>. Modern medical research and chemical laboratory analysis show that a chemical substance named as ginsenoside is contained in the Radix Ginseng and has the obvious effects of regulating the central nervous system of human, strengthening heart, resisting fatigue, regulating substance metabolism and the like, and therefore a good therapeutic effect is played on the multiple diseases of the nervous system, the cardiovascular system, the endocrine system and reproductive system.
The Radix Ginseng can regulate the balance of the excitation process and inhibition process of the central nervous system. The results of research of the influence on brain electrical activities of animals of the Radix Ginseng show that: the Radix Ginseng influences the two nerve processes of excitation and inhibition, but mainly strengthen the excitation process of the cerebral cortex. Due to the fact that the ginseng acts on the inhibition process at the same time, the inhibition tends to concentration, and differentiation is accelerated and is more complete. The Radix Ginseng can regulate nerve functions and enable the disordered nerve process caused by tension to be restored.
Some reports suggest that Radix Ginseng extract has an antagonism effect on poor memory caused by anisodine and pentobarbital sodium, can also improve memory consolidation impairment caused by cycloheximide and sodium nitrite and memory reappearance deficits caused by 40% ethyl alcohol. Radix Ginseng stem-leaf saponins are injected into the abdominal cavity of rats in dosages of 200 mg/kg, 100 mg/kg and 50 mg/kg, the anisodine effect can be obviously resisted, the memory of the rats can be obviously improved, intracerebral RNA can be obviously increased, but no obvious effect is produced on the DNA and protein content.
The Radix Ginseng also has obvious impact on cerebral blood flow and cerebral energy metabolism. A Radix Ginseng preparation can improve glucose intake of the rabbit brain, at the same time, reduces the ratio of lactic acid, pyruvic acid and lactic acid/pyruvic acid, and makes the utilization of glucose to transfer to aerobic metabolism from an anaerobic metabolism pathway. The Radix Ginseng also can make free inorganic phosphorus in the cerebral cortex increase by 25%. The Radix Ginseng fruit saponin can improve oxygen uptake ability of the brain. The total Radix Ginseng saponin and total Radix Ginseng root saponin both have protection against cerebral ischemia/reperfusion injury. In sum, the Radix Ginseng can make animal brains more reasonably utilize energy substance glucose, generates energy with oxidation, and synthesizes more ATP for activity use such as learning, memorizing and the like.
The ginsenoside Rg3 is a Radix Ginseng rare saponin, which is only contained in radix ginseng rubra over more than 5 years for 3 parts per 10 million, and is the most essential component in the Radix Ginseng. The ginsenoside Rg3 has improving and preventing effects on various diseases, and has a great therapeutic effect on elderly common diseases, such as cardiovascular and cerebrovascular diseases, coronary heart disease, limbs acratia, disability, memory impairment.
The ginsenoside Rg3 is Radix Ginseng glycol tetracyclic triterpenoid saponin, it is found in practical applications that since the ginsenoside Rg3 has a larger molecular weight (785.02), its lipid solubility and water solubility are both relatively poor, making its oral absorption percentage being about 10.28%, and resulting in relatively poor absorption of oral preparations, results of animal pharmacokinetic experiments indicate that about 80% of the medicine in native form is not absorbed and is excreted through feces, and the medicine is not fully used in the body, which will influence full use of its clinical therapeutic effect. Therefore, the ginsenoside Rg3 is made into topical preparations, which directly enters the lymphatic system by absorption via neck skin and posterior auricular mucosa, it is absorbed via the blood-brain barrier so as to achieve a therapeutic effect on the cerebral nervous system, and a better clinical therapeutic effect is obtained and further confirmed through pharmacological experiments and clinical applications.
Borneolum Syntheticum: studies of immunohistochemical staining technique adopted by Zhao Baosheng, et al. show that Borneolum Syntheticum can not induce expression of ICAM-1 on rat brain microvascular EC, but can significantly reduce adhesion of leukocytes and EC during brain trauma, and has a protective effect on EC, the studies prove that the promotion of openness of the blood-brain barrier with borneolum syntheticum is irrelevant to ICAM-1, and borneolum syntheticum has a certain protective effect on the rat brain tissue with brain trauma. Rats are divided into 12 groups by Ge Chaoli, et al., drenched with a borneolum syntheticum paraffin oil solution 10, 13 mL·kg, and are also divided into 1, 2, 4, 8, 24, 48 hours for each group according to the difference of sampling time points, the control group is drenched with equal amount of paraffin oil solution. The two groups of rats are sacrificed and brains are taken at above-mentioned time points, respectively, and observed with a transmission electron microscope. The studies show that borneolum syntheticum has a significant influence on tight junction between rat cerebral capillary endothelial cells; compared with the control group, intercellular tight junction gaps are widened, are discontinous, and are reduced in structure, after drench with borneolum syntheticum for 4 hours, changes start to occur, reach to peak at the 8th hour, and return to normal at the 48th hour, which indicates that borneolum syntheticum can make ultra microstructures of the rat blood-brain barrier have reversible changes. The rabbit serum containing traditional Chinese medicine borneolum syntheticum prepared by Chen Yanming uses verapamil as positive control medicine, it is observed and found with an MTT method that on MDCK (Madin-Darby Canine Kidney) and HeLa (human uterine cancer) cell lines, borneolum syntheticum can both significantly increase cell toxicity caused by vincristine, and has a similar effect with verapamil, which illustrates that borneolum syntheticum has a significant inhibition effect on expression of P-glycoprotein. There are also studies which show that promotion of physiological openness of the blood-brain barrier with borneolum syntheticum is due to the fact that after borneolum syntheticum enters the blood-brain barrier, affinity of borneolum syntheticum with P-glycoprotein is large, as a result, the borneolum syntheticum is pumped out of cells, thus the vascular active substance 5-HT which has small affinity with P-glycoprotein is accumulated in cells so as to promote openness of the blood-brain barrier.
Styrax: rats are divided into a sulpiride combined storax group and a single-use sulpiride group, after the rats continuously take oral medication for one week, the surgery for installing probes in the brain and neck of rats are performed, the equilibration is waited for one hour, dialysate in the blood of right atrium of rats and dialysate in intracerebral hippocampal tissues are collected at each time period (30, 60, 90, 120, 150, 180 minutes) by blood microdialysis and brain microdialysis methods, respectively, the content of sulpiride in the sample is detected by a reversed-phase high-performance liquid fluorescence chromatogram system, and the content of sulpiride in the brain and the blood are compared after medicine administration by a statistic method. As a result, it is obvious that the concentrations of sulpiride in the brain and blood of rats in the sulpiride combined storax group are higher than that in the single-use sulpiride group, the concentration ratio of sulpiride in the blood and brain of rats is 1:0.2 in the single-use sulpiride group, the concentration ratio of sulpiride in the blood and brain of rats is raised to 1:0.3 in the sulpiride combined storax group, and compared with the single-use sulpiride group, the concentration of sulpiride in the brain is raised by 39% and the concentration of sulpiride in the blood is raised by 69% in the sulpiride combined storax group. This indicates that the storax can significantly increase the concentrations of sulpiride in the brain and blood of rats, and the storax can facilitate the sulpiride to penetrate the gastrointestinal barrier and the blood-brain barrier.
Moschus: expert experiments indicate that: a Moschus aqueous solution or suspension which is used for intravenous injection by 50 mg/kg or for lateral ventricle injection by 2.5 mg/kg can de-synchronize the quiet and awake rabbit cortex electroencephalogram (EEG) for a short time, and part of animals are with restlessness behavior in a watch state, indicating that the cerebral cortex can be excited and the cortical electrical activity can be enhanced; the Moschus aqueous solution has an obvious wake-up effect on anesthetized rabbits, lateral ventricle injection is more effective than intravenous injection, and the above specification may describe that the Moschus likely penetrates the blood-brain barrier to directly act on the central nervous system.
Rhizoma Acori Tatarinowii: rhizoma acori tatarinowii is one of traditional resuscitation-inducing aromatic herbs, is singly used among the people or is compatible with other traditional Chinese medicines for wide usage, is mainly used for treating encephalopathy and has a definite curative effect, and it prompted that the rhizoma acori tatarinowii has a certain effect on the BBB. Volatile oil is extracted by adopting the volatile oil extraction method I described in the appendix I of the Chinese Pharmacopoeia 2005, the extraction rate is about 1.58%, and an oil removing water decoction is concentrated to be 1 g/ml. A gas chromatography-mass spectrometer (GC/MS) is adopted to detect that a volatile oil sample contains α-asarone, β-asarone, guaiene and acorenone, the cerebrospinal fluid of a rat taking the volatile oil is detected under the same conditions, and a result shows that the α-asarone and the β-asarone can penetrate the BBB to enter the cerebrospinal fluid. A liquid chromatography-mass spectrometer (HPLC/MS) is adopted to determine an oil removing water decoction sample and the cerebrospinal fluid sample of a rat drenched with the oil removing water decoction, three compounds are detected, the molecular weights are 396, 452 and 339 respectively, and a known rhizoma acori tatarinowii compound is not yet included therein. The ultra microstructures of the cerebral cortex BBB of the rats before and after medicine administration are observed through the transmission electron microscope, it can be found that all extracted parts of the rhizoma acori tatarinowii enable the tight junction (TJ) of BBB endothelial cells to be loose, but the BBB is basically complete and is not yet destroyed. Borneolum syntheticum is used as a positive medicine to preliminarily discuss the promoting effect on BBB passage of Evans blue (EB) of the extracted parts of the rhizoma acori tatarinowii, namely the EB content in the brain reflects the permeability of the BBB, the statistical analysis shows that the EB content in the brain is remarkably higher than that of a blank group (P<0.01) 2.5 hours after the borneolum syntheticum is filled into the stomach, and the self-administrating medicine with the rhizoma acori tatarinowii dose of 11.7 g/kg does not produce an obvious effect within 0.5 to 2.5 hours; the administration time of the rhizoma acori tatarinowii is prolonged, the EB content in the brain after 5 hours is remarkably higher than that of the blank group (P<0.05) when the volatile oil doses of the rhizoma acori tatarinowii are 23.4 g/kg and 11.7 g/kg, and a dose dependency trend exists; when the total dose is 11.7 g/kg, the EB content in the brain after 5 hours is remarkably higher than that of the blank group (P<0.01). One week after the extracted parts of the rhizoma acori tatarinowii are filled into the stomachs of the rats, phenytoin sodium is injected into the abdominal cavity, an HPLC method is adopted to detect the concentration of the phenytoin sodium in the brain, and a result shows that: the content of the phenytoin sodium in the brain of the total-group animal taking the rhizoma acori tatarinowii is higher than that in the brain of the animal group which does not take the rhizoma acori tatarinowii, but a higher concentration (P<0.01) is still kept after 4 hours. An HPLC-ECD is utilized to detect the concentration of 5-hydroxytryptamine (5-HT) in brain tissues, and statistical analysis shows that: compared with the blank group, the content of the 5-HT in the brain of the rat in the volatile oil group is increased (P<0.05) and the content of the 5-HT in the rhizoma acori tatarinowii water decoction group and the content of 5-HT in the total medicine group are remarkably increased (P<0.01). A HeLa cell line is applied to serve as a carrier for evaluating the activity of the rhizoma acori tatarinowii to P-glycoprotein on the BBB, and the parts (volatile oil, water decoction and total medicine) of the rhizoma acori tatarinowii can obviously strengthen the cytotoxicity of vincristine (VCR) after 0.5 hour (P<0.01); the volatile oil part can still obviously strengthen the cytotoxicity of the VCR after 8 hours (P<0.01), and the medicine effect maintaining time of the volatile oil is obviously longer than that of the positive medicine verapamil (VER) (P<0.01); the positive medicine and the parts of the rhizoma acori tatarinowii have no obvious effect on the cytotoxicity of the VCR after 24 hours (P>0.05). In sum, the rhizoma acori tatarinowii can affect the ultra microstructure of the rat BBB, strengthen the permeability of the BBB to the EB and promote increase of the content of the CNS medicine phenytoin sodium entering the brain and has the pharmacological actions of activating the BBB and leading medicines to the brain. Action mechanism research results show that the 5-HT content regulation by the rhizoma acori tatarinowii is one of its BBB permeability increase mechanisms, and in addition, the rhizoma acori tatarinowii has a very strong effect of inhibiting exocytosis of the P-glycoprotein medicine on the HeLa cell membranes, which is also an action mechanism of BBB permeability increase.
Radix Curcumae: the curcumin contained in the Radix Curcumae has an effect of inhibiting the senile dementia. The main cause of the dementia is intertwining of amyloid β. The curcumin enables the intertwined amyloid β to be loosened. The curcumin is a powerful antioxidant and anti-inflammatory chemical molecule and can penetrate the strictly guarded blood-brain barrier to neutralize oxides and copper ions having oxidative toxicity, and beyond that, the curcumin can further inhibit a cell transcription factor (NF kappa B) causing inflammation. Therefore, the curcumin is a key ingredient for protecting the brain tissues and decreasing dementia. In India, the number of patients with dementia is only ten percent of those in other countries, and curcumin's contributions cannot go unnoticed.